Event Title

Finding optimal OPA-IBLC Derivatized Amino Acid Analysis Parameters with 3-dimensional Fluorometric Scans for HPLC Method Validation

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Session

Department of Chemistry & Biochemistry

Event Website

https://www.andrews.edu/services/research/research_events/conferences/urs_honors_poster_symposium/index.html

Start Date

3-26-2021 1:40 PM

End Date

3-26-2021 2:00 PM

Department

Chemistry and Biochemistry

Description

This research validates RP-HPLC fluorimeter and OPA-IBLC derivatization parameters to lower limits of detection (LOD) and quantitation (LOQ). A 3-dimensional fluorometric scan in pH 10.4 sodium borate buffer showed peak OPA-IBLC derivatized histidine and glutamic acid fluorescence occurs at 368- to 445-nm and 336- to 452-nm λ¬ex-λ¬em, respectively. HPLC-FLD response was refined with systematic FLD analysis in pH 4 sodium acetate buffer after column separation. Derivatization reaction completion was ensured by increasing injector mixing time until peak integration area stabilized. Results inform future FLD parameters and determine racemic OPA-IBLC derivatized amino acid ratios in ostrich eggshells.

Acknowledgments

Advisor: Lisa Ahlberg

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Mar 26th, 1:40 PM Mar 26th, 2:00 PM

Finding optimal OPA-IBLC Derivatized Amino Acid Analysis Parameters with 3-dimensional Fluorometric Scans for HPLC Method Validation

This research validates RP-HPLC fluorimeter and OPA-IBLC derivatization parameters to lower limits of detection (LOD) and quantitation (LOQ). A 3-dimensional fluorometric scan in pH 10.4 sodium borate buffer showed peak OPA-IBLC derivatized histidine and glutamic acid fluorescence occurs at 368- to 445-nm and 336- to 452-nm λ¬ex-λ¬em, respectively. HPLC-FLD response was refined with systematic FLD analysis in pH 4 sodium acetate buffer after column separation. Derivatization reaction completion was ensured by increasing injector mixing time until peak integration area stabilized. Results inform future FLD parameters and determine racemic OPA-IBLC derivatized amino acid ratios in ostrich eggshells.

https://digitalcommons.andrews.edu/honors-undergraduate-poster-symposium/2021/symposium/7